Preparation method and application of chromone compound

Abstract

The invention discloses a preparation method and application of chromone compounds used for Alzheimer's disease drugs, and the chromone compounds are shown as any one of 1)-5): 1) (E)-6-bromo-2-[(2-(6-bromobenzo [d] [1, 3] dioxane-5-yl) vinyl)]-4H-chromene-4-ketone 2) (E)-2, 3, 4-triazole-5-yl) vinyl]-4H-chromene-4-ketone, 2) (E)-6-bromo-2-[(2-(6-bromobenzo [d] [1, 3] dioxane-5-yl) vinyl]-4H-chromene-4-ketone; the invention discloses a chromone compound and a preparation method of the chromone compound, and the chromone compound is 1, 4-dihydroxy-N '-[(6-hydroxy-4-oxo-4H-chromene-3-yl) methylene] phenylhydrazone 3) (3E, 3' E)-3, 3 '-((1, 4-phenylenediamine) bis (methylene alkenyl)) bis (6-hydroxy-2-methoxychromene-4-ketone). The compound disclosed by the invention has a relatively good acetylcholin esterase inhibiting effect, shows a good effect of inhibiting Abeta42 polypeptide aggregation and promoting Abeta42 fiber degradation, can avoid more toxic and side effects, and improves the medication safety. The method has a good application prospect in screening of anti-senile dementia drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of drug synthesis, and relates to the synthesis, preparation method and application of chromone compounds. Background Art

[0002] Alzheimer's disease is a multi-cause systemic degenerative disease of the cerebral cortex that often occurs in the elderly. Recent memory impairment is the main clinical symptom of early Alzheimer's disease, and the concentration of acetylcholine in the brain is believed to be related to recent memory.

[0003] Early studies have found that the pathogenesis of Alzheimer's disease is the cholinergic hypothesis. Cholinergic neurons can synthesize a large amount of acetylcholine, which is transported to the cerebral cortex and hippocampus through projection fibers. In the patient's cerebrospinal fluid and brain tissue, the severe loss, degeneration and functional defects of cholinergic neurons damage the acetylcholinergic nerves, leading to learning and memory impairment and cognitive impairment, and producing Alzheimer's symptoms. This suggests that intervention with cholinesterase inhibitors may play a role in treating Alzheimer's disease.

[0004] Cholinesterase inhibitors are currently the most commonly used cholinergic transmitter-related drugs, such as galantamine, huperzine A, rivastigmine, etc. Chromone drug components in other natural products have antioxidant, free radical scavenging, and central cholinergic function enhancement effects. They can selectively enhance the effects of acetylcholine in the cerebral cortex and hippocampus, significantly improve patients' cognitive functions and daily living abilities, and significantly alleviate the severity of the disease.

[0005] Chromone compounds are a class of widely existing and biologically active substances. By modifying the structure of different positions of chromone compounds, the purpose of acetylcholine inhibition can be achieved. It is expected that more highly effective and low-toxic drugs will be developed in the future and benefit mankind. Summary of the invention

[0006] In view of the above problems, the present invention provides a chromone compound for preparing a drug for Alzheimer's disease, a preparation method thereof and use thereof. The compound has good safety and can avoid more toxic side effects.

[0007] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0008] The present invention provides use of a chromone compound for preparing a drug for Alzheimer's disease.

[0009] Furthermore, the chromone compound is any one of the following:

[0010] (E)-6-bromo-2-[(2-(6-bromobenzo[d][1,3]dioxetyl-5-yl)vinyl)]-4H-chromen-4-one, referred to as A1, has the structural formula:

[0011] (E)-2,4-dihydroxy-N'-[(6-hydroxy-4-oxo-4H-chromen-3-yl)methylene]benzohydrazone, referred to as A2, has the structural formula:

[0012] (3E,3'E)-3,3'-((1,4-phenylenediamino)bis(methylenevinyl))bis(6-hydroxy-2-methoxychromene-4-one), referred to as A3, has the structural formula:

[0015] The present invention also provides a method for preparing the chromone compounds for the above-mentioned purpose, comprising the following steps:

[0016] S1: 2-hydroxy-5-bromoacetophenone, ethyl acetate, sodium hydride and tetrahydrofuran are mixed into a solution, refluxed, and after the reaction is completed, the solvent is evaporated to dryness to obtain a solid, which is then extracted with dichloromethane, washed with glacial acetic acid, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the first product;

[0017] S2: The first product is mixed with glacial acetic acid and concentrated sulfuric acid, and refluxed. After the reaction is completed, the mixture is cooled to room temperature; distilled water is added and allowed to stand overnight; the mixture is taken out and filtered, and placed in an oven overnight to obtain the second product;

[0018] S3: Mix 6-bromo-3,4-methylenedioxybenzaldehyde with the second product, add methanol and sodium methoxide solution, reflux, and cool to room temperature after the reaction is completed; add distilled water and let stand overnight; take out the mixed solution and filter it, and place it in an oven overnight to obtain the target product;

[0019] S4: After the reaction is completed, extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, suction filtration, concentration, and drying are performed in sequence; and chromatography separation is performed to obtain the target product A1;

[0020] S5: phosphorus oxychloride and N,N-dimethylamide are mixed into a solution, and refluxed for reaction. After the reaction is completed, the solution is allowed to stand overnight to obtain a first product.

[0021] S6: Mix the first product with 2,5-dihydroxyacetophenone, and reflux at 10°C and normal pressure. After the reaction is completed, evaporate the solvent to obtain a solid, then extract with dichloromethane, wash with glacial acetic acid, wash with saturated brine, dry with anhydrous sodium sulfate, filter and concentrate to obtain a second product;

[0022] S7: mixing the second product in S6 with 2,4-dihydroxybenzohydrazide and methanol, and subjecting the mixture to a reflux reaction at room temperature and pressure overnight. After the reaction is completed, cooling the mixture to room temperature to obtain the target product;

[0023] S8: After the reaction is completed, extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, suction filtration, concentration, and drying are performed in sequence; and chromatography separation is performed to obtain the target product A2;

[0024] S9: mixing the second product in S6 with p-phenylenediamine and methanol solution, and subjecting the mixture to a reflux reaction at room temperature and pressure overnight. After the reaction is completed, cooling the mixture to room temperature to obtain the target product; obtaining the target product;

[0025] S10: After the reaction is completed, extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, suction filtration, concentration, and drying are performed in sequence; and chromatography separation is performed to obtain the target product A3;

[0026] Furthermore, in step S1, the molar ratio of sodium hydride to tetrahydrofuran is 1:1.

[0027] Furthermore, in step S1, the reflux temperature is 80° C. and the time is 3 hours.

[0028] Furthermore, in steps S2 and S3, the reflux temperature is 100° C. and the time is 4 hours.

[0029] Furthermore, the concentration of glacial acetic acid used for washing was 10%.

[0030] Furthermore, in step S2, the volume ratio of glacial acetic acid to concentrated sulfuric acid is 100:1.

[0031] Further, in step S3, the concentration of the sodium ethoxide solution is 20%.

[0032] Further, in step S6, stirring is carried out at room temperature overnight.

[0033] The present invention has the following beneficial effects:

[0034] The present invention innovatively synthesizes a chromone compound, explores its safety, and proves that it has low toxicity.

[0035] The invention cleverly designs a method for preparing chromone compounds.

[0036] The compound prepared by the invention has good acetylcholinesterase inhibition effect and low toxicity, thereby improving drug safety. The invention provides experimental basis and theoretical foundation for the research and development of novel chromone derivatives. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 The inhibitory effect of compound A1 in Example 1 of the present invention on the aggregation of Aβ42 polypeptide

[0038] Figure 2 The inhibitory effect of compound A2 of Example 1 of the present invention on the aggregation of Aβ42 polypeptide

[0039] Figure 3 The inhibitory effect of compound A3 in Example 1 of the present invention on the aggregation of Aβ42 polypeptide

[0040] Figure 4 The effect of compound A1 in Example 1 of the present invention on promoting the degradation of Aβ42 fibers

[0041] Figure 5 The effect of compound A2 in Example 1 of the present invention on promoting the degradation of Aβ42 fibers

[0042] Figure 6 The effect of compound A3 in Example 1 of the present invention on promoting the degradation of Aβ42 fibers

[0043] Figure 7 The hydrogen nuclear magnetic resonance spectrum of compound A1 in Example 1 of the present invention is

[0044] Figure 8 The carbon NMR spectrum of compound A1 of Example 1 of the present invention is

[0045] Fig. 9 The mass spectrum of compound A1 of Example 1 of the present invention is

[0046] Fig.10 The nuclear magnetic resonance hydrogen spectrum of compound A2 in Example 1 of the present invention is

[0047] Fig.11 The carbon NMR spectrum of compound A2 in Example 1 of the present invention is

[0048] Fig.12 The mass spectrum of compound A2 of Example 1 of the present invention is

[0049] Fig.13 The hydrogen nuclear magnetic resonance spectrum of compound A3 in Example 1 of the present invention is

[0050] Fig.14 The carbon NMR spectrum of compound A3 of Example 1 of the present invention is

[0051] Fig.15 The mass spectrum of compound A3 of Example 1 of the present invention is DETAILED DESCRIPTION

[0052] In order to facilitate a better understanding of the present invention, the following examples are used to illustrate the present invention. These examples belong to the protection scope of the present invention, but do not limit the protection scope of the present invention.

[0053] Example 1

[0054] Its synthetic route is as follows: Compound A1: Compound A2: Compound A3:

[0055] Product A1, white solid, yield: 74%, melting point 177.2-197.0℃, 1 H NMR (400MHz, DMSO-d6) δ8.07 (t, J=2.4Hz, 1H), 7.94 (dt, J=8.9, 2.7Hz, 1H), 7.58 (d, J=8.9H z, 1H), 7.14 (d, J=5.2Hz, 2H), 6.33-6.14 (m, 2H), 6.07 (dd, J=9.9, 1.0Hz, 2H), 5.86 (s, 1H). 13 C NMR (101MHz, DMSO) δ175.94, 167.49, 155.35, 147.96, 147.90, 137.17, 137.01, 127.4 3, 125.14, 121.45, 118.13, 112.26, 111.60, 107.81, 102.41, 69.72.HRMS(m / z)[M+H] + : calculated forC18H10Br2O4, 450.8998; found, 450.8911.

[0056] Product A2, white solid, yield: 79%, melting point 233.2-234.2℃, 1 H NMR (400MHz, DMSO-d6) δ12.41 (s, 1H), 11.79 (s, 1H), 10.21 (d, J = 33.3Hz, 2H), 8.77 (s, 1H), 8.61 (s, 1H), 7.82 (d, J = 8.8Hz, 1H), 7.61 (dd, J=9.1, 5.2Hz, 1H), 7.40 (dd, J=4.6, 2.9Hz, 1H), 7.29 (dt, J=9.0, 2.7Hz, 1H), 6.37 (dd, J=8.8, 2.4Hz, 1H), 6.31 (d, J=2.4Hz, 1H). 13C NMR (101MHz, DMSO) δ175.31, 175.17, 166.13, 163.29, 163.14, 156.79, 156.00, 155.82, 154.75, 150.00, 14 1.45, 129.97, 124.91, 124.75, 124.00, 120.59, 117.62, 108.20, 107.89, 106.29, 103.33.HRMS(m / z)[M+Na] + : calculated for C17H12N2O6, 363.0588; found, 363.0581.

[0057] Product A3 is a light yellow powder solid with a yield of 55% and a melting point of 317.6-318.5°C. 1 H NMR (400MHz, DMSO-d6) δ11.89 (d, J=12.6Hz, 2H), 9.37 (s, 2H), 8.13-8.02 (m, 2H ), 7.43(s, 4H), 7.20-7.17(m, 2H), 6.96-6.90(m, 4H), 5.73(s, 2H), 3.35(s, 6H). 13 C NMR (101MHz, DMSO) δ180.30, 152.52, 148.64, 145.37, 136.43, 123.58, 122.3 8, 119.23, 118.38, 110.56, 103.89, 101.39, 55.11, 54.96.HRMS(m / z)[M+Na] + : calculated for C28H24N2O8, 517.1605; found, 517.1460.

[0058] Acetylcholinesterase inhibition test results

[0059] The following biological test examples are described to illustrate the present invention.

[0060] 1. The inhibitory activity of the compounds of the present invention can be determined by using the following assay system:

[0061] Select acetylcholinesterase and prepare it to 3U / ml, and DTNB to 1mmol / L. Add 150μl PBS, 20μl AchE, 50μl DTNB, 50μl ATch, and 30μl sample diluent to the 96-well plate in sequence. After mixing and shaking, place it at 37°C for five minutes and detect it with an enzyme marker. Measure the absorbance (A) value of each well, according to the formula: Inhibition rate = (average A value of the negative control group - average A value of the drug group) / (average A value of the control group - A value of the blank control group) * 100%.

[0062] Experimental results on inhibiting Aβ42 peptide aggregation and promoting Aβ42 fiber degradation

[0063] The following biological test examples are described to illustrate the present invention.

[0064] 1. The inhibitory activity of the compounds of the present invention can be determined by using the following assay system:

[0065] Aβ42 sample preparation: Aβ42 film was dissolved in 69.5 μl of a solution prepared with 300 μM CH3CN, 250 mM Na2CO3 and NaOH (48.3:48.3:3.4, v / v / v), and the mixture was dissolved by brief sonication and vortexing. The obtained alkaline Aβ42 solution (500 μM) was diluted 1:10 with phosphate buffer (10 mM, pH 7.7) containing NaCl (11 mM) to obtain a 50 μM Aβ42 solution with a final pH of 8.0 and a NaCl concentration of 10 mM. The final assay conditions were as follows: 50 μM Aβ42 in phosphate buffer (8.7 mM) containing 10 mM NaCl, 14.5 μM Na2CO3, 0.85 mM NaOH and 8.2% acetonitrile (final pH 8.0). The Aβ42 solution was sonicated briefly and incubated at 30 °C without any agitation and then analyzed at the selected time.

[0066] Aggregation assay of Aβ42 polypeptide: Compound A1 was dissolved in methanol with a stock concentration of 2.0 mM and diluted in assay buffer. Compounds A2 and A3 were dissolved and diluted in the same way. The concentrations were measured (0, 0.625, 1.25, 2.5, 5, 10 μM). In the time course experiment, different concentrations of the stock solution of chromone compounds (A1, A2, A3) were added to the Aβ42 polypeptide sample, and then incubated at 30°C and detected every 30 minutes using an ELISA reader.

[0067] Decomposition of Aβ42 fibers: Compound A1 was dissolved in methanol with a stock concentration of 2.0 mM and diluted in the assay buffer. Compounds A2 and A3 were dissolved and diluted in the same way. The concentrations were measured (0, 2.5, 10 μM). In the time course experiment, different concentrations of the stock solution of chromone compounds (A1, A2, A3) were added to the Aβ42 fiber samples, then incubated at 30°C and detected every 30 minutes using an ELISA reader.

[0068] The results are shown in Table 1

[0069] Table 1 Acetylcholinesterase inhibition test results Compound Inhibitory activity IC50 (μmol / L) A1 2.52μmol / L A2 5.56μmol / L A3 1.78μmol / L

[0070] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of chromone compounds for treating Alzheimer's disease.

2. The use of the chromone compound according to claim 1 for preparing a drug for treating Alzheimer's disease, characterized in that: The chromone compound is any one of the following: [0010] (E)-6-bromo-2-[(2-(6-bromobenzo[d][1,3]dioxetyl-5-yl)vinyl)]-4H-chromene-4-one, referred to as A1, has the structural formula: [0011] (E)-2,4-dihydroxy-N′-[(6-hydroxy-4-oxo-4H-chromen-3-yl)methylene]benzohydrazone, referred to as A2, has the structural formula: [0012](3E,3′E)-3,3′-((1,4-phenylenediamino)bis(methylene olefin))bis(6-hydroxy-2-methoxychromene-4-one), referred to as A3, has the structural formula:

3. A method for preparing a chromone compound for use as claimed in claim 1 or 2, characterized in that: The following steps are involved: S1: 2-hydroxy-5-bromoacetophenone, ethyl acetate, sodium hydride and tetrahydrofuran are mixed into a solution, refluxed, and after the reaction is completed, the solvent is evaporated to dryness to obtain a solid, which is then extracted with dichloromethane, washed with glacial acetic acid, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the first product; S2: The first product is mixed with glacial acetic acid and concentrated sulfuric acid, and refluxed. After the reaction is completed, the mixture is cooled to room temperature; distilled water is added and allowed to stand overnight; the mixture is taken out and filtered, and placed in an oven overnight to obtain the second product; S3: Mix 6-bromo-3,4-methylenedioxybenzaldehyde with the second product, add methanol and sodium methoxide solution, reflux, and cool to room temperature after the reaction is completed; add distilled water and let stand overnight; take out the mixed solution and filter it, and place it in an oven overnight to obtain the target product; S4: After the reaction is completed, extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, suction filtration, concentration, and drying are performed in sequence; and chromatography separation is performed to obtain the target product A1; S5: phosphorus oxychloride and N,N-dimethylamide are mixed into a solution, and refluxed for reaction. After the reaction is completed, the solution is allowed to stand overnight to obtain a first product. S6: Mix the first product with 2,5-dihydroxyacetophenone, and reflux at 10°C and normal pressure. After the reaction is completed, evaporate the solvent to obtain a solid, extract it with dichloromethane, wash it with glacial acetic acid, wash it with saturated brine, dry it with anhydrous sodium sulfate, filter it with suction, and concentrate it to obtain the second product; S7: mixing the second product in S6 with 2,4-dihydroxybenzohydrazide and methanol, and subjecting the mixture to a reflux reaction at room temperature and pressure overnight. After the reaction is completed, cooling the mixture to room temperature to obtain the target product; S8: After the reaction is completed, extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, suction filtration, concentration, and drying are performed in sequence; and chromatography separation is performed to obtain the target product A2; S9: mixing the second product in S6 with p-phenylenediamine and methanol solution, and subjecting the mixture to a reflux reaction at room temperature and pressure overnight. After the reaction is completed, cooling the mixture to room temperature to obtain the target product; Obtaining the target product; S10: After the reaction is completed, extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, suction filtration, concentration and drying are performed in sequence; and chromatography separation is performed to obtain the target product A3.

4. The method for preparing chromone compounds according to claim 3, characterized in that: In step S1, the molar ratio of sodium hydride to tetrahydrofuran is 1:

1.

5. The method for preparing chromone compounds according to claim 3, characterized in that: In step S1, reflux is performed at room temperature and pressure for 12 hours.

6. The method for preparing chromone compounds according to claim 3, characterized in that: In steps S2 and S3, the reflux temperature is 100° C. and the time is 4 hours.

7. The method for preparing chromone compounds according to claim 3, characterized in that: In step S2, the volume ratio of glacial acetic acid to concentrated sulfuric acid is 100:

1.

8. The method for preparing chromone compounds according to claim 3, characterized in that: In steps S2 and S3, the oven static temperature is 65°C.

9. The method for preparing chromone compounds according to claim 3, characterized in that: Step S6, temperature 10°C, time 12 hours.

10. The method for preparing chromone compounds according to claim 3, characterized in that: Step S7, normal temperature and pressure, time is 12 hours.

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